I have my vector and insert double digests purified and gel extracted. After ligating and transforming, I didn't observe any colonies on the plate. Is it okay if I use the purified double digests for ligation again? or Do i have to do a new double digestion?
Yes it is fine to store. Usually they are kept in the fridge until you confirm your final plasmid. Alternatively, some people like to keep a large digested stock of plasmid they constantly use frozen (like a CAS9 plasmid while constantly swapping the guide) to speed up the work flow.
Hi Hanna, You suggest to store keep this double digest in -20deg?
I did not get transformed colonies post ligation using these double digests. Do you suggest using these to set up another ligation?
Since you do not know which part of the ligation is causing the problem I would suggest to keep them and try again since I assume you were able to see the double cut band when you purified it.
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